Inflammation Mediators Regulate the Microbiota Resistance to Adverse Factors.

We studied the effects of IL-1ß, IL-8, TNFα, and prostaglandin E2α in concentrations typically observed in health and during inflammation on the growth of vaginal microbiota and its resistance to factors inhibiting the synthesis of proteins, nucleic acids, and peptidoglycans. An increase in the cytokine levels, characteristic of inflammation, inhibits the growth of Lactobacillus population and improves its resistance to adverse factors. The growth of the population of opportunistic microorganisms (S. aureus, E. coli) is stimulated under these conditions, while their resistance to adverse factors decreases. Hence, it seems that the cytokines regulate the behavior of the host cells and of its bacterial symbionts.

Proinflammatory Cytokines as Regulators of Vaginal Microbiota.

It was shown that IL-1ß, IL-8, and IL-6 in concentrations similar to those in the vagina of healthy women stimulated the growth of normal microflora (Lactobacillus spp.) and suppressed the growth and biofilm production by S. aureus and E. coli. On the contrary, these cytokines in higher concentrations typical of vaginal dysbiosis suppressed normal microflora and stimulated the growth of opportunistic microorganisms. TGF-ß1 in both doses produced a stimulating effects on study vaginal microsymbionts. It is hypothesized that pro-inflammatory cytokines serve as the molecules of interspecies communication coordinating the interactions of all components of the vaginal symbiotic system.

[EFFECT OF MICROSYMBIONTS ON THE ABILITY OF VAGINAL EPITHELIOCYTES TO MODIFY BIOLOGICAL PROPERTIES OF BACTERIA].

AIM: Study regularities of effects of various types of vaginal microsymbionts on the ability of mucosal epitheliocytes of the vagina to modify biological properties of bacteria. MATERIALS AND METHODS: Effect of thermo-inactivated cells of Staphylococcus aureus, Escherichia coli, H2O2-producing and H2O2-non-producing lactobacilli on the ability of primary vaginal epitheliocytes to alter growth and antagonistic activity of Staphylococcus aureus, Escherichia colt, H2O2-producing and H2O2-nonproducing lactobacilli was studied using a multi-component module system. RESULTS: Alterations of composition of vaginal epitlieliocyte exometabolites under the effect of S. aureus and E. coli was established to result in a pronounced stimulation of antagonistic activity of H2O2-producing and an increase of the number of H2O2-nonproducing lactobacilli. Thermo-inactivated cells of lactobacilli stimulated production of metabolites by epitheliocytes, that suppress the growth and antagonistic activity of allochthonous microflora and stimulate similar parameters of lactobacilli. The strongest effects on the ability of vaginal epitheliocytes to alter biological properties of bacteria were rendered by H2O2-producing lactobacilli. CONCLUSION: The regularities identified allow to examine epitheliocytes and normoflora of vagina as a symbiotic system, the coordinated interaction of its components is directed on maintaining microecological stability of female reproductive tract.

[Effect of IL-1beta on growth properties of vaginal microsymbionts].

AIM: Study the effect of IL-1beta in concentrations that are characteristic for vaginal normo- and pathocenosis on growth properties of vaginal microsymbionts. MATERIALS AND METHODS: Concentration of IL-1beta in vaginal contents of women during bacterial vaginosis and normocenosis was determined by using enzume immunoassay. Changes of growth characteristics and biofilm formation ability of Staphylococcus aureus, Escherichia coli, Lactobacilus spp., Corynebacterium spp. under the effect of various IL-1beta concentrations by method of O'Toole G.A. (1999) were studied. RESULTS: IL-1beta in concentrations characteristic for normocenosis was shown to be able to cause stimulating effect on growth properties of lactobacilli and corynebacteria and suppress growth of S. aureus and E. coli in both plankton and biofilm cultures. IL-1beta concentrations characteristic for vaginal dysbiosis on the contrary result in suppression of growth of lactobacilli biomass against the background of stimulation of growth properties and biofilm formation ability of S. aureus and E. coli. CONCLUSION: Differential dose-dependent effect of IL-1beta on biomass growth and biofilm formation ability of vaginal microsymbionts is a mechanism of regulation of vaginal microbiocenosis.

[Role of dominant microflora in mechanisms of woman vaginal biotope protection].

AIM: Detection of properties of the dominant microsymbiont emerging in the course of interaction of associative symbiosis components and ensuring protection of vaginal mucosa from seeding by pathogens. MATERIALS AND METHODS: H202-producing Lactobacillus spp., Corynebacterium spp., Staphylococcus aureus, Escherichia coli and primary epitheliocytes from lower reproductive tract of women were used. The ability of Lactobacillus spp. to influence changes of synthesis of catalase inhibitors by corynebacteria, bactericidal activity of lysozyme, lactoferrin and thrombocyte antimicrobial protein, antimicrobial activity of primary vaginal epitheliocytes was evaluated. Changes of antagonistic and growth properties oflactobacilli during interaction with secretory products of Corynebacterium spp., S. aureus, E. coli and vaginal epitheliocytes were also studied. RESULTS: Exometabolites of both epitheliocytes and corynebacteria were revealed to stimulate in most cases lactobacilli biomass growth and Sincreased their antagonistic activity against S. aureus u E. coli. Metabolites of H202-producing lactobacilli increased synthesis of antimicrobial substances by epitheliocytes and potentiated bactericidity of natural resistance factors. Under the influence of lactobacilli metabolites an increase of production of catalase inhibitors by corynebacteria was revealed. CONCLUSION: Symbiotic interrelations of dominant microorganisms with host organism and associants under the condition of associative symbiosis leading to stimulation of production and potentiating of the effect of antibacterial protection factors are the basis for colonization resistance of vaginal biotope.

[Production of cytokines by vaginal epitheliocytes in the process of interaction with dominant and associative microsymbionts].

AIM: Study the expression of cytokines by vaginal epitheliocytes in the process of interaction with dominant and associative microsymbionts. MATERIALS AND METHODS: IL-8, IL-6, IL-1beta and TNFalpha expression in response to interaction with heat inactivated Lactobacillus spp., Staphylococcus aureus, Escherichia coli, Corynebacterium spp. or their secretory products in comparison with basal expression of cytokines by vaginal epitheliocytes was studied. Results. Lactobacilli secretory products were shown not to influence the expression of IL-8 and IL-1beta and moderately stimulated IL-6 and TNFalpha expression. Contact of epitheliocytes with heat inactivated lactobacilli increased secretion of IL-8, IL-6 and IL-1beta and reduced TNFalpha production. Secretory products of S. aureus and E. coli caused stimulation of IL-6, IL-1beta production and practically did not change the expression of IL-8 and TNFalpha. Contact of epitheliocytes with heat inactivated S. aureus sup pressed TNFalpha production and had no influence on IL-8, IL-6 and IL-1beta expression, contact with E. colistimulated TNFalpha and IL-1beta expression and suppressed IL-6 expression. Changes in cytokine expression during interaction of epitheliocytes with corynebacteria were largely similar to the results of interaction with lactobacilli except IL-6 production that was markedly stimulated by corynebacteria secretory products. Conclusion. In epithelial-bacterial interactions dominant and associative microorganisms have a differential effect on functional status of mucosal epitheliocytes manifesting in production of cytokines that could be the basis of mucosal immunity regulation.

[Features of epithelial-bacterial interactions in bacterial vaginosis].

AIM: Study of features of interactions of mucosal epitheliocytes of vagina with dominant and associative vaginal microsymbionts in bacterial vaginosis. MATERIALS AND METHODS: Changes in growth characteristics and ability to form biofilms of Staphylococcus aureus, Escherichia coli, Lactobacillus spp. under the influence of secretory products of vaginal epitheliocytes obtained from 36 women with bacterial vaginosis and 32 practically healthy women were studied. RESULTS: Vaginal epitheliocytes of women with bacterial vaginosis were shown to change their functional activity during contact with microorganisms and their secretory products, causing differential influence on the properties of symbiont bacteria. A suppression of growth of dominant microsymbionts--lactobacilli, mostly their peroxide producing biovars, and a weakening of suppressing effect on the growth of biomass of associative microorganisms accompanied by stimulation of their ability to form biofilms was noted. Preliminary contact of an epitheliocyte with bacterial cell or its supernatant increased the intensity of the changes of properties of bacteria under the influence of secretory products of epitheliocytes. CONCLUSION: Suppression of growth of lactobacilli (mostly their peroxide producing biovars) and stimulation of the ability to for biofilms of the associative microorganisms as a result of bacterial-epithelial interactions in bacterial vaginosis determine mechanisms of formation and stability of pathomicrobiocenosis and a reduction of colonization resistance of vaginal biotope.

[Role of associative microsymbionts in functioning of associative symbiosis].

AIM: Study the role of associative microsymbionts in biocenosis based on comparative evaluation of interbacterial and epithelial-bacterial interactions on the example of vaginal biotope. MATERIALS AND METHODS: Corynebacterium spp., Staphylococcus aureus, Escherichia coli and Lactobacillus spp. and primary epitheliocytes isolated from the lower part of reproductive tract of women were used. Interactions by associant-macropartner vector were evaluated by changes of growth properties and antagonistic activity of the associant under the influence of exometabolites of vaginal epitheliocytes and expression of cytokines by epitheliocytes under the influence of associants. The nature of interaction by associant-dominant vector was evaluated by changes of adhesive properties and biofilm formation of associants and bactericidal activity of peroxide-producing lactobacilli. RESULTS: Groups of associants that are opposite by their function in symbiosis were isolated. An example of associants that stabilize associative symbiosis by stimulating growth and antagonistic activity of dominant, moderate increase of cytokine production by epitheliocytes are corynebacteria, whose growth, biofilm formation, adhesion and antagonism during interaction with dominant and macropartner are also increased. The nature of interaction of associants of the other group (S. aureus and E. coli) with dominant and macropartner is the opposite. The determinative role of hydrogen peroxide in mechanisms of differentiation of associative microsymbionts and functioning of associative symbiosis ofvagina was shown. CONCLUSION: The proposed approach allows to differentiate associative microsymbionts with various ecological roles, evaluate their contribution into maintenance of the stability of the symbiosis and open new possibilities of management of quality of microbiocenosis.

[Epithelial-bacterial interactions as a basis of microbiocenosis formation].

AIM: Evaluate significance of epithelial-bacterial interactions for the formation and/or support of microbiocenoses. MATERIALS AND METHODS: Effect of vaginal epitheliocytes on growth properties of lactobacilli strains obtained from the same biotope from 16 women with vaginal normocenosis was studied. During intravaginal probiotic therapy in 24 women with vaginal dysbiosis probability of colonization of vagina by probiotic strain was evaluated depending on the result of effect of vaginal epitheliocytes on its growth properties. RESULTS: Exometabolites of epitheliocytes in normocenosis were shown to render stimulating effect on the growth of autostrain of dominant microflora, that probably is the basis of formation and/or support of microbiocenosis. Stimulating effect of exometabolites of vaginal epitheliocytes of patients on growth properties of probiotic strain was revealed to be the prerequisite for successful probiotic therapy. CONCLUSION: The data obtained allow to examine epithelial-bacterial interactions as a basis of formation of microbiocenoses. Evaluation of these interactions may be used for individual selection of probiotic strains.

[Influence of microbial-epithelial interactions on biological characteristics of vaginal microflora].

AIM: Evaluation of influence of microbial epithelial interactions on growth properties and antagonistic activity of dominant and associated vaginal microflora. MATERIALS AND METHODS: Growth characteristics and antagonistic activity changes were studied in associated (Staphylococcus aureus, Escherichia coli, Corynebacterium spp.) and dominant (Lactobacilus spp.) microflora during contact with vaginal epitheliocytes and influence of vaginal epitheliocyte secretory products by using the developed technique. RESULTS: Secretory products of vaginal epitheliocytes and vaginal epitheliocytes differentially influence growth factors of vaginal microbiocenosis. Suppression of S. aureus and E. coli biomass growth based predominately on extracellular secretory products of epitheliocytes was observed. Stimulation of growth properties was prevalent for lactobacilli and corynebacteria and was observed in contact interaction as well as during effect of epitheliocyte extracellular secretory products. CONCLUSION: The resulting bacterial-epithelial interaction is a differentiating feature between autochthonous and allochthonous microflora due to the creation of selective advantages for dominant microsymbionts by growth properties and antagonistic activity stimulation, and biomass growth suppression of associated symbionts.

[The microflora of the reproductive tract in women during intrauterine interventions]. / Mikroflora reproduktivnogo trakta zhenshchin pri vnutrimatochnykh vmeshatel'stvakh.

The species compositions and persistence factors of the vaginal and cervical microflora of the reproductive tract of women in cases of intrauterine interventions (medical abortion, intrauterine contraception) were studied. Women with inflammatory complications following intrauterine interventions were found to have the same species of bacteria in their vaginal and cervical microflora. In addition, an increase in the values of the persistence factors of vaginal microflora was registered in women practicing intrauterine contraception and a decrease in the persistence potential of vaginal microflora was registered after abortion.

[The correction of dysbiosis of the female reproductive tract by using an estradiol-containing preparation]. / Korrektsiia disbioza reproduktivnogo trakta zhenshchin s pomoshch'iu éstriolsoderzhashchego preparata.

The influence of Ovestin, an estradiol-containing preparation, on the vaginal microflora in women of the reproductive age in dysbiosis was studied. As the result of the local application of this preparation, the normalization of microflora was noted in 92% of women, which was manifested by an increase in the number of lactobacteria and indigenous corynebacteria, a decrease in the number, or elimination, of opportunistic microorganisms and the suppression of their capacity for persistence.

[Microbiological approaches to the prognostication of inflammatory complications after intrauterine interventions]. / Mikrobiologicheskie podkhody k prognozirovaniiu vospalitel'nykh oslozhnenii vnutrimatochnykh vmeshatel'stv.

The species composition and persistence parameters of vaginal microflora in women after intrauterine interventions (intrauterine contraception and artificial abortion) has been studied. The relationship between inflammatory complications after intrauterine interventions and previous changes in the microflora has been established. Criteria for the evaluation of vaginal microbiocenosis, making it possible to prognosticate the appearance of inflammatory complications after intrauterine interventions, have been worked out.

[The characteristics of the vaginal microflora in intrauterine contraception]. / Kharakteristika vaginal'noi mikroflory pri vnutrimatochnoi kontratseptsii.

The results of the study of vaginal microbiocenosis in women using intrauterine contraceptives are presented. Inflammatory complications in intrauterine contraception were shown to be linked with vaginal dysbiosis. Disturbances in vaginal microbiocenosis were characterized by the deficiency of lactoflora and the presence of enterobacteria, Staphylococcus aureus, Gardnerella of fungi of the genus Candida. The problem of the possibility of complications of microbial etiology in women using intrauterine contraceptives are discussed.